JP2021191234A - Cell culture tank - Google Patents

Abstract

To provide a cell culture tank that can stabilize the pressure acting on cellular tissue in the cell culture tank.SOLUTION: A cell culture tank (50) has a first chamber (51) having a first opening (55) formed at its top, a liquid feed pipe (42) to feed culture medium (L) into the first chamber, and cellular tissue (B) disposed inside the first chamber at a position with a predetermined depth downward from the first opening. The culture medium is continuously fed from the liquid feed pipe into the first chamber so that the culture medium continuously pours from the first opening.SELECTED DRAWING: Figure 2

Description

The present invention relates to a tank for culturing cells.

Conventionally, cell culture in which a pressurized gas is supplied into a tank containing a culture solution by a pressurizing means, and the pressurizing means is controlled so that the flow rate of a flow meter provided between the tank and the culture device becomes a predetermined flow rate. There is a system (see Patent Document 1).

Japanese Unexamined Patent Publication No. 2005-168436

By the way, in the cell culture system described in Patent Document 1, the pressurizing pressure by the pressurized gas is controlled so that the flow rate of the culture solution becomes a predetermined flow rate. However, in culturing a cell tissue having blood vessels, it is desirable that the pressure acting on the cell tissue is stable from the viewpoint of the pressure resistance of the blood vessels and the expression of biological functions. In the cell culture system described in Patent Document 1, the pressure acting on the cell tissue in the culture apparatus (cell culture tank) cannot be stabilized.

The present invention has been made to solve the above problems, and a main object thereof is to provide a cell culture tank capable of stabilizing the pressure acting on the cell tissue in the cell culture tank.

The first means for solving the above-mentioned problems is a cell culture tank.
The first room with the first opening formed at the top,
A liquid supply pipe that sends the culture solution to the inside of the first chamber,
Inside the first chamber, a cell tissue arranged at a predetermined depth downward from the first opening,
Equipped with
The culture solution is continuously supplied from the liquid supply pipe to the inside of the first chamber so that the culture solution continuously overflows from the first opening.

According to the above configuration, a first opening is formed in the upper part of the first chamber. The liquid feeding pipe sends the culture liquid to the inside of the first chamber. Therefore, if the culture solution is continuously sent from the liquid supply pipe to the inside of the first chamber, the culture solution overflows from the first opening of the first chamber.

Here, inside the first chamber, the cell tissue is arranged at a position at a predetermined depth downward from the first opening. Then, the culture liquid is continuously sent from the liquid feeding pipe to the inside of the first chamber so that the culture liquid continuously overflows from the first opening. When the culture solution continuously overflows from the first opening, the depth of the culture solution inside the first chamber is constant. Therefore, the cell tissue is always arranged at a predetermined depth from the first opening in the culture medium. Therefore, the hydraulic pressure of the culture solution acting on the cell tissue can be made constant, and the pressure acting on the cell tissue in the cell culture tank can be stabilized.

The second means is a cell culture tank,
The first room with the first opening formed at the top,
A liquid supply pipe that sends the culture solution to the inside of the first chamber,
Inside the first chamber, a cell tissue arranged at a predetermined depth downward from the first opening,
Equipped with
The culture solution is sent from the liquid feeding pipe to the inside of the first chamber in the predetermined cycle so that the culture solution overflows from the first opening in a predetermined cycle.

According to the above configuration, the culture solution is sent from the liquid feeding pipe to the inside of the first chamber at a predetermined cycle so that the culture solution overflows from the first opening at a predetermined cycle. In the state where the culture solution overflows from the first opening or immediately after the overflow, the depth of the culture solution inside the first chamber is constant. Therefore, the cell tissue is arranged at a predetermined depth from the first opening in the culture medium. Therefore, the hydraulic pressure of the culture solution acting on the cell tissue can be made constant, and the pressure acting on the cell tissue in the cell culture tank can be stabilized.

In the third means, the second opening is formed at a position lower than the first opening in the upper part, the culture solution overflowing from the first opening flows in, and the culture solution overflows from the second opening. It includes a second chamber and a sensor that is inserted into the second chamber and measures the properties of the culture solution stored inside the second chamber.

Since the properties of the culture medium change depending on the activity of the cell tissue, it is desirable to measure the properties of the culture medium constantly or regularly. On the other hand, when measuring the properties of the culture solution, it is necessary not to contaminate the culture solution.

In this respect, according to the above configuration, the second opening is formed at a position lower than the first opening in the upper part of the second chamber. The culture solution overflowing from the first opening of the first chamber flows into the second chamber, and the culture solution overflows from the second opening of the second chamber. Then, the sensor is inserted into the inside of the second chamber and measures the properties of the culture solution stored in the inside of the second chamber.

Therefore, the properties of the culture medium stored inside the second chamber can be maintained at the same level as the properties of the culture medium stored inside the first chamber, and the culture medium changed by the activity of the cell tissue can be maintained. The properties can be measured by a sensor. Further, the culture solution does not flow from the first chamber to the second chamber, and the culture solution does not flow back from the second chamber to the first chamber. Therefore, while making it possible to measure the properties of the culture solution, it is possible to prevent the culture solution in the first chamber from being contaminated when measuring the properties of the culture solution. Moreover, since the sensor is inserted inside the second chamber, it is possible to prevent the sensor from interfering with the maintenance of the first chamber.

In the fourth means, the first chamber and the second chamber are adjacent to each other via a first wall extending in the vertical direction, and the upper end of the first wall forms a part of the first opening. A third chamber adjacent to the second chamber is provided via a second wall extending in the vertical direction, and the upper end of the second wall forms a part of the second opening. The upper end of the wall is lower than the upper end of the first wall.

According to the above configuration, the first chamber and the second chamber are adjacent to each other via the first wall extending in the vertical direction. The upper end of the first wall forms part of the first opening. Therefore, the culture solution overflowing from the first chamber beyond the upper end of the first wall flows into the second chamber. The third chamber is adjacent to the second chamber via the second wall extending in the vertical direction. The upper end of the second wall forms part of the second opening, and the upper end of the second wall is lower than the upper end of the first wall. Therefore, the culture solution overflowing from the second chamber beyond the upper end of the second wall flows into the third chamber, and the culture solution does not flow back from the second chamber to the first chamber. Therefore, while making it possible to measure the properties of the culture solution, it is possible to prevent the culture solution in the first chamber from being contaminated when measuring the properties of the culture solution.

In the fifth means, a first lid covering the upper part of the first room, a second lid covering the upper part of the second room and the third room, and a roof portion covering the upper part of the first wall are provided. The sensor is inserted into the inside of the second chamber through the second lid, and the end of the first lid on the second chamber side and the end of the second lid on the first chamber side. The end portion is arranged on the roof portion.

It is desirable that the first to third rooms are provided with a lid to prevent germs and foreign substances from entering. However, it is necessary to open and close the lid of the first chamber in order to perform regular maintenance of the cell tissue. On the other hand, when the sensor penetrates the lid of the second chamber and is inserted into the inside of the second chamber, it is desirable to suppress the opening and closing of the second lid. For that purpose, it is desirable that the lid of the first chamber and the lid of the second chamber can be opened and closed separately. In the fourth means described above, the first chamber and the second chamber are adjacent to each other via the first wall extending in the vertical direction. Therefore, there is a possibility that germs and foreign substances may fall from the gap between the lid of the first chamber and the lid of the second chamber into the first chamber and the second chamber.

In this regard, according to the above configuration, the cell culture tank includes a first lid covering the upper part of the first chamber and a second lid covering the upper part of the second chamber and the third chamber. Therefore, it is possible to prevent germs and foreign substances from entering the first to third chambers. Further, in the configuration in which the sensor penetrates the second lid and is inserted into the inside of the second chamber, only the first lid can be opened and closed.

Further, the cell culture tank is provided with a roof portion that covers the upper part of the first wall. The end of the first lid on the second chamber side and the end of the second lid on the first chamber side are arranged on the roof portion. Therefore, even if germs or foreign substances fall from the gap between the first lid and the second lid, they can be received by the roof portion, and it is possible to prevent the germs and foreign substances from entering the first and second chambers. can.

The sixth means includes a main body that surrounds the second chamber and the third chamber together with the first wall, and the second lid and the main body have the second chamber and the upper part of the third chamber above the third chamber. A first engagement that engages with each other so as to restrict the vertical movement of the second lid with respect to the main body by moving the second lid in a direction away from the first chamber after covering with the two lids. A portion and a second engaging portion are provided, respectively, and the upper part of the first chamber is covered with the first lid in a state where the first engaging portion and the second engaging portion are engaged with each other. Therefore, it is restricted to move the second lid in a direction closer to the first chamber.

In a configuration in which the sensor penetrates the second lid and is inserted into the inside of the second chamber, the second lid may unintentionally open due to the sensor tilting or something hitting the sensor.

In this regard, according to the above configuration, the cell culture tank includes a main body that surrounds the second chamber and the third chamber together with the first wall. That is, the second room and the third room are surrounded by the first wall and the main body. For the second lid and the main body, after covering the upper part of the second chamber and the third chamber with the second lid, the second lid is moved in the direction away from the first chamber, so that the second lid is in the vertical direction with respect to the main body. A first engaging portion and a second engaging portion that engage with each other are provided so as to restrict the movement of the two. Therefore, by engaging the first engaging portion and the second engaging portion, it is possible to restrict the vertical movement of the second lid with respect to the main body, and it is possible to prevent the second lid from unintentionally opening. can do.

Further, by covering the upper part of the first chamber with the first lid in a state where the first engaging portion and the second engaging portion are engaged, the second lid is moved in the direction of approaching the first chamber. Is regulated. Therefore, by covering the upper part of the first chamber with the first lid, it is possible to prevent the first engaging portion from being disengaged from the second engaging portion, and the second lid is unintentionally opened. That can be further suppressed.

In the seventh means, the first lid is formed with an introduction port for introducing gas into the first chamber. Therefore, a gas for adjusting the properties of the culture solution stored in the first chamber can be introduced into the inside of the first chamber from the introduction port. Further, the gas introduced into the inside of the first chamber from the introduction port is introduced into the second chamber through the first opening and is introduced into the third chamber through the second opening. Therefore, the properties of the culture solution in the second chamber can be brought close to the properties of the culture solution in the first chamber.

In the eighth means, the first lid is formed with an introduction port for introducing gas into the first chamber, and the second lid has a portion of the main body surrounding the third chamber. A thin portion is formed in which the gap between the second lid and the second lid is wider than the gap between the portion of the main body surrounding the second chamber and the second lid. Therefore, the gas introduced into the third chamber is easily discharged to the outside of the third chamber through the gap between the main body and the thin-walled portion, and the gas is easily discharged from the first chamber to the second chamber and from the second chamber to the third chamber in order. It becomes easier to guide the gas. Therefore, the properties of the culture broth in the second chamber can be further brought closer to the properties of the culture broth in the first chamber.

Schematic diagram showing a cell culture system. The schematic diagram which shows the operation of the cell culture system when the pump is stopped. The schematic diagram which shows the operation of the cell culture system at the time of driving a pump. Perspective view of the cell culture tank. Perspective view of the main body of the cell culture tank. Top view of the main body of the cell culture tank. Side view of the main body of the cell culture tank. Partial cross-sectional perspective view of the main body of the cell culture tank. FIG. 7 is a sectional view taken along line IX-IX in FIG. A perspective view of the second cover. Bottom view of the second cover. Assembly drawing of DO sensor and pH sensor. The cross-sectional view which shows the modification example of the 1st partition wall.

Hereinafter, an embodiment embodied in a cell culture tank of a cell culture system for culturing cells will be described with reference to the drawings.

As shown in FIG. 1, the cell culture system 10 includes a pressure-controlled liquid feeding unit 20, a cell culture tank 50, a pump 90, and the like.

The pressure control liquid feeding unit 20 includes an air supply source 21, a flow rate proportional valve 24, a throttle valve 27, a sterilization filter 31, a tank 34, a pressure sensor 37, a liquid level sensor 41, a control unit 45, and the like.

The air supply source 21 (gas supply source) supplies air (gas) at a predetermined pressure. The air supply source 21 is connected to the tank 34 by pipes 22 and 23. The pipe 22 is provided with a flow rate proportional valve 24. The flow rate proportional valve 24 (flow rate adjusting unit) can adjust the opening degree in proportion to the input signal, and adjusts the flow rate of the air supplied from the air supply source 21. The flow rate proportional valve 24 is controlled by the control unit 45.

The pipe 22 and the pipe 23 are connected at the connection portion 25. The pipe 23 is provided with a sterility filter 31. The sterilization filter 31 sterilizes the air flowing to the tank 34 through the pipe 23. The tank 34 is formed in a bottomed cylindrical shape, is airtight, and stores the culture solution L. The pipe 23 is connected to the upper part of the tank 34. That is, the pipe 23 is connected to the tank 34 at a position higher than the liquid level of the culture liquid L and is not in contact with the culture liquid L. The first pipe is composed of the pipes 22 and 23. The shape of the tank 34 is not limited to the bottomed cylindrical shape, and is arbitrary.

A reagent that is free in the pH range of 6 to 8 and has a pH buffering function, such as bicarbonate, is added to the culture solution L. As the bicarbonate, sodium bicarbonate (sodium hydrogencarbonate), potassium hydrogencarbonate, calcium hydrogencarbonate and the like can be adopted. In addition, serum, amino acids, vitamins, sugars and the like are added to the culture solution L as nutrients necessary for the activity of cell tissue B (cells) having blood vessels. An indicator such as phenol red is added to the culture solution L in order to visually confirm the pH.

The pressure sensor 37 detects the pressure of the culture solution L at the lower part of the tank 34. The liquid level sensor 41 detects the position of the liquid level of the culture solution L.

A pipe 26 is connected to the connection portion 25. The connection portion 25 is connected to the air discharge portion 28 by a pipe 26. The throttle valve 27 is provided in the pipe 26. The throttle valve 27 (throttle portion) is, for example, a needle valve that adjusts a minute flow rate, and limits (adjusts) the flow rate of air discharged to the discharge section 28. The opening degree of the throttle valve 27 is set to a predetermined opening degree so that the pressure of the air in the tank 34 (the pressure in the tank 34) becomes the target pressure. The opening degree of the throttle valve 27 may be changed according to the target pressure of the air in the tank 34. The air flowing to the discharge unit 28 is discharged to the outside (atmosphere) of the pressure control liquid supply unit 20. The second pipe is composed of the pipes 23 and 26.

The cell culture tank 50 includes a culture chamber 51, a sensor chamber 52, and a regulation chamber 53. The culture chamber 51 (first chamber) and the sensor chamber 52 (second chamber) are adjacent to each other via a first partition wall 61 (first wall) extending in the vertical direction. The sensor chamber 52 and the adjustment chamber 53 (third chamber) are adjacent to each other via a second partition wall 62 (second wall) extending in the vertical direction. The upper end 62a of the second partition wall 62 is lower than the upper end 61a of the first partition wall 61. Details of the culture chamber 51, the sensor chamber 52, the adjustment chamber 53, the first partition wall 61, and the second partition wall 62 will be described later.

The liquid feeding pipe 42 connects the inside of the culture liquid L stored in the tank 34 to the culture chamber 51 of the cell culture tank 50. The liquid feeding pipe 42 is inserted into the culture liquid L stored in the tank 34, and the lower end of the liquid feeding pipe 42 is located at a position lower than the liquid level of the culture liquid L.

The cell culture tank 50 stores the culture solution L and the cell tissue B. The culture solution L is sent from the tank 34 to the cell culture tank 50 via the liquid supply pipe 42. The cell tissue B is, for example, an animal cell tissue having blood vessels. The cell tissue B is cultured in the culture medium L and expresses a predetermined biological function. At that time, the dissolved oxygen concentration DO and pH of the culture solution L in the cell culture tank 50 change due to the activity of the cells.

The culture solution L is sent to the culture chamber 51 of the cell culture tank 50 at a pressure (pressurization) of 10 [mmHg] or less so as not to damage the blood vessels of the cell tissue B. Further, in order for the cell tissue B to exhibit a predetermined biological function, a pressure-controlled liquid is used to send the culture liquid L at the target pressure, not a flow-controlled liquid to send the culture liquid L at the target flow rate. Is desirable. The reason for this is that in an actual animal cell tissue, blood is pumped by the heart, which is closer to a pressure-controlled fluid than a flow-controlled fluid. Further, in order for the cell tissue B to express a predetermined biological function, it is desirable that the pressure acting on the cell tissue B is stable. When the culture solution L is sent to the cell culture tank 50 at a pressure (pressurization) of 10 [mmHg] or less, the flow rate of the culture solution L is 1 [mL / min] or less.

An oxygen concentration sensor 81 and a pH sensor 82 (see FIG. 4) are inserted in the culture solution L stored in the sensor chamber 52. The oxygen concentration sensor 81 (sensor) detects the dissolved oxygen concentration DO (property) of the culture solution L in the sensor chamber 52. The pH sensor 82 (sensor) detects the pH (property) of the culture solution L in the sensor chamber 52. When the pH of the culture solution L is detected by the pH sensor 82, the reference solution (internal solution) may leak slightly from the pH sensor 82 to the culture solution L.

The adjustment chamber 53 of the cell culture tank 50 is connected to the tank 34 by a liquid return pipe 43. The return liquid pipe 43 (supply pipe) is provided with a pump 90 and a check valve 92. The pump 90 pumps the culture solution L when the position of the liquid level detected by the liquid level sensor 41 is lower than the predetermined position (liquid level sensor 41: OFF). Then, the pump 90 stops the pumping of the culture solution L when the position of the liquid level detected by the liquid level sensor 41 rises above the predetermined position (liquid level sensor 41: ON). The check valve 92 (check valve) allows the flow of the culture solution L from the pump 90 to the tank 34, and prohibits the flow of the culture solution L from the tank 34 to the pump 90. The liquid return pipe 43 is connected to the tank 34 at a position lower than the liquid level of the culture liquid L and higher than the lower end of the liquid feed pipe 42.

The control unit 45 is a microcomputer including a CPU, ROM, RAM, an input / output interface, and the like. The control unit 45 controls the flow rate proportional valve 24 so as to send the culture solution L to the cell culture tank 50 at the target pressure.

When the culture liquid L is sent from the tank 34 to the cell culture tank 50 via the liquid delivery pipe 42, the control unit 45 has a flow rate proportional valve 24 so that the target flow rate of air calculated based on predetermined conditions is supplied. To control. The control unit 45 maintains the state for a predetermined time of 5 [S] or less (for example, 3 [S]) until the pressure Pv in the tank 34 is in equilibrium. In this embodiment, the pressure Pv is in equilibrium at about 1 [S]. As a result, the culture solution L is sent to the cell culture tank 50 at the target pressure.

The time from the start of supplying air from the air supply source 21 to the equilibration of the pressure Pv inside the tank 34 is that the larger the volume of the tank 34, the larger the effective cross-sectional area S of the throttle valve 27, the larger the time of the tank 34. The higher the internal target pressure, the longer it will be. In this respect, in the culture of the cell tissue B having blood vessels, it is necessary to send the culture solution L to the cell culture tank 50 at a slight pressure of 10 [mmHg] or less, so that the target pressure inside the tank 34 is very low. It is set. Therefore, the time until the pressure Pv equilibrates becomes very short.

The control unit 45 is supplied from the air supply source 21 in a state where the pressure inside the tank 34 is balanced when the culture liquid L is sent from the tank 34 to the cell culture tank 50 via the liquid feed pipe 42. The tank is controlled with the flow rate proportional valve 24 so that the target flow rate of air corresponding to the target pressure inside the tank 34 is supplied based on a predetermined correlation between the flow rate of air and the pressure inside the tank 34. Equilibrate the pressure inside 34. In other words, the control unit 45 is based on a predetermined correlation between the flow rate of the air supplied from the air supply source 21 in a state where the pressure inside the tank 34 is balanced and the pressure inside the tank 34. In a state where the flow rate proportional valve 24 is controlled so that the air of the target flow rate corresponding to the target pressure inside the 34 is supplied, the pressure inside the tank 34 is balanced, and the culture is performed from the tank 34 via the liquid feed pipe 42. The liquid L is sent to the cell culture tank 50.

FIG. 2 is a schematic diagram showing the operation of the cell culture system 10 when the pump 90 is stopped.

The pressure inside the tank 34 is controlled to the target pressure, and the culture solution L in the tank 34 is pressurized. Therefore, the culture solution L is sent from the tank 34 to the culture chamber 51 of the cell culture tank 50 via the liquid supply pipe 42. Along with this, the liquid level of the culture solution L in the tank 34 is lowered. The culture solution L in the culture chamber 51 is filled up to the upper end 61a of the first partition wall 61. Then, the culture solution L in the culture chamber 51 overflows to the sensor chamber 52 beyond the upper end 61a of the first partition wall 61.

Inside the culture chamber 51 for storing the culture solution L, the cell tissue B is arranged at a predetermined depth D downward from the bottom surface of the culture chamber 51, that is, the upper end 61a of the first partition wall 61. Then, the culture liquid L is continuously sent from the liquid feeding pipe 42 to the inside of the culture chamber 51 so that the culture liquid L continuously overflows beyond the upper end 61a of the first partition wall 61. In a state where the culture solution L continuously overflows beyond the upper end 61a of the first partition wall 61, the depth of the culture solution L inside the culture chamber 51 is constant. Therefore, the cell tissue B is always arranged at a predetermined depth D from the upper end 61a of the first partition wall 61 in the culture medium L.

The dissolved oxygen concentration DO of the culture solution L in the sensor chamber 52 is detected by the oxygen concentration sensor 81. The pH of the culture solution L in the sensor chamber 52 is detected by the pH sensor 82. The culture solution L in the sensor chamber 52 is filled up to the upper end 62a of the second partition wall 62. Then, the culture solution L in the sensor chamber 52 overflows to the adjusting chamber 53 beyond the upper end 62a of the second partition wall 62. Along with this, the liquid level of the culture solution L in the adjustment chamber 53 is rising.

Then, when the position of the liquid level detected by the liquid level sensor 41 is lower than the predetermined position (liquid level sensor 41: OFF), the pump 90 starts driving and pumps the culture liquid L.

FIG. 3 is a schematic diagram showing the operation of the cell culture system 10 when the pump 90 is driven.

Even when the pump 90 is driven, the pressure inside the tank 34 is controlled to the target pressure, and the culture solution L in the tank 34 is pressurized. Therefore, the culture solution L is sent from the tank 34 to the culture chamber 51 of the cell culture tank 50 via the liquid supply pipe 42. The flow rate of the culture solution L pumped from the adjustment chamber 53 to the tank 34 by the pump 90 is larger than the flow rate of the culture solution L sent from the tank 34 to the culture chamber 51 by the pressurized air. Therefore, the liquid level of the culture solution L in the tank 34 is rising. The culture solution L in the culture chamber 51 is filled up to the upper end 61a of the first partition wall 61. Then, the culture solution L in the culture chamber 51 overflows to the sensor chamber 52 beyond the upper end 61a of the first partition wall 61.

The dissolved oxygen concentration DO of the culture solution L in the sensor chamber 52 is detected by the oxygen concentration sensor 81. The pH of the culture solution L in the sensor chamber 52 is detected by the pH sensor 82. The culture solution L in the sensor chamber 52 is filled up to the upper end 62a of the second partition wall 62. Then, the culture solution L in the sensor chamber 52 overflows to the adjusting chamber 53 beyond the upper end 62a of the second partition wall 62. The flow rate of the culture solution L pumped from the adjustment chamber 53 to the tank 34 by the pump 90 is larger than the flow rate of the culture solution L overflowing from the sensor chamber 52 to the adjustment chamber 53. Therefore, the liquid level of the culture solution L in the adjustment chamber 53 is lowered.

Then, when the position of the liquid level detected by the liquid level sensor 41 rises above the predetermined position (liquid level sensor 41: ON), the pump 90 stops driving and stops the pumping of the culture liquid L. After that, the cell culture system 10 repeats from the operation of FIG.

Next, the configuration of the cell culture tank 50 will be described in detail. FIG. 4 is a perspective view of the cell culture tank 50. The cell culture tank 50 includes a main body 60, a first cover 75, a second cover 78, an oxygen concentration sensor 81, a pH sensor 82, a stirrer 86, and the like.

FIG. 5 is a perspective view of the main body 60 of the cell culture tank 50, and FIG. 6 is a plan view of the main body 60 of the cell culture tank 50. The main body 60 is formed of a hollow rectangular parallelepiped shape of metal, resin, or the like. The upper surface of the main body 60 is open. The culture chamber 51, the sensor chamber 52, and the adjustment chamber 53 are formed inside the main body 60.

The first cover 75 (first lid) is formed of metal, resin, or the like, and covers the upper part of the culture chamber 51. The first cover 75 is formed with an introduction port 76 for introducing gas into the inside of the culture chamber 51. Oxygen or carbon dioxide can be adopted as the gas to be introduced from the introduction port 76. A tube (piping) or the like is connected to the introduction port 76, and oxygen and carbon dioxide are introduced into the culture chamber 51 through the tube. The second cover 78 (second lid) is formed of metal, resin, or the like, and covers the upper part of the sensor chamber 52 and the adjustment chamber 53. The oxygen concentration sensor 81 and the pH sensor 82 penetrate the second cover 78 and are inserted into the sensor chamber 52. The details of the second cover 78 will be described later.

A stirrer 86 is arranged below the culture chamber 51. The stirrer 86 uses a magnetic force to rotate a stirrer (not shown) to stir the culture solution L in the culture chamber 51. The stirrer is arranged in the culture chamber 51 so as not to hit the cell tissue B. The stirrer and the cell tissue B may be displaced in the horizontal direction or may be displaced in the vertical direction.

The liquid feeding pipe 42 is connected to the culture chamber 51 via a joint 83. The liquid return pipe 43 is connected to the adjusting chamber 53 via a joint 84.

7 is a side view of the main body 60 of the cell culture tank 50, FIG. 8 is a partial cross-sectional perspective view of the main body 60 of the cell culture tank 50, and FIG. 9 is a sectional view taken along line IX-IX of FIG.

As shown in FIG. 8, the culture chamber 51 and the sensor chamber 52 are adjacent to each other via the first partition wall 61. The first partition wall 61 is formed in a rectangular plate shape by metal, resin, or the like. The first partition wall 61 extends vertically upward from the bottom surface of the main body 60. The first partition wall 61 extends over the entire length of the culture chamber 51, and horizontally partitions the culture chamber 51 and the sensor chamber 52. The culture solution L in the culture chamber 51 can flow into the sensor chamber 52 only from above the first partition wall 61, and cannot flow into the sensor chamber 52 from the side of the first partition wall 61. The culture chamber 51 is surrounded by the first partition wall 61 and the main body 60.

At the upper part of the culture chamber 51, a first opening 55 having a rectangular cross section that opens in the horizontal direction is formed. The upper end 61a of the first partition wall 61 forms the bottom (part) of the first opening 55. It can also be said that an opening (first opening) having a rectangular cross section that opens upward is formed in the upper part of the culture chamber 51.

As shown in FIGS. 8 and 9, the sensor chamber 52 and the adjusting chamber 53 are adjacent to each other via the second partition wall 62. The second partition wall 62 is formed in the shape of a rectangular plate by metal, resin, or the like. The second partition wall 62 extends vertically upward from the bottom surface of the main body 60. The second partition wall 62 extends over the entire width of the sensor chamber 52 and horizontally partitions the sensor chamber 52 and the adjustment chamber 53. The culture solution L in the sensor chamber 52 can flow into the adjusting chamber 53 only from above the second partition wall 62, and cannot flow into the adjusting chamber 53 from the side of the second partition wall 62. The sensor chamber 52 and the adjustment chamber 53 are surrounded by the first partition wall 61 and the main body 60.

A second opening 56 having a rectangular cross section that opens upward is formed in the upper part of the sensor chamber 52. The upper end 62a of the second partition wall 62 forms a measuring portion (part) of the second opening 56. The upper end 62a of the second partition wall 62 is lower than the upper end 61a of the first partition wall 61. That is, the second opening 56 is formed at a position lower than that of the first opening 55. It can also be said that an opening (second opening) having a rectangular cross section that opens in the horizontal direction is formed in the upper part of the sensor chamber 52.

As shown in FIGS. 1, 6 to 8, the upper part of the first partition wall 61 is covered with the roof portion 65. The roof portion 65 is formed to have a width wider than the thickness of the first partition wall 61, and extends horizontally along the upper end 61a of the first partition wall 61. That is, the roof portion 65 covers the upper portion of the end portion of the culture chamber 51 on the sensor chamber 52 side and the upper portion of the end portion of the sensor chamber 52 on the culture chamber 51 side. The lower surface of the roof portion 65 forms the upper surface (part) of the first opening 55.

At the end of the roof portion 65 on the culture chamber 51 side, a rectangular plate-shaped side plate 66 extending upward is provided. The side plate 66 extends to the upper end of the main body 60. The side plate 66 is provided over the entire length of the culture chamber 51. At the end of the roof portion 65 on the sensor chamber 52 (adjustment chamber 53) side, a rectangular plate-shaped side plate 67 extending upward is provided. The side plate 67 extends to the upper end of the main body 60. The side plate 67 is provided over the entire length including the sensor chamber 52 and the adjustment chamber 53.

As shown in FIGS. 5 to 8, "L" -shaped grooves 68A and 68B are formed in the outer edge portion of the upper part of the main body 60 in a side view. The groove 68A (second engaging portion) and the groove 68B (second engaging portion) are formed in the main body 60 so as to face each other at a position closer to the sensor chamber 52 than the culture chamber 51. The grooves 68A and 68B extend downward from the upper end of the main body 60 and then horizontally extend in a direction away from the culture chamber 51. The grooves 68A and 68B are formed with a predetermined width W2 and a predetermined depth D2.

As shown in FIGS. broken. When the culture chamber 51 is covered with the first cover 75, the outer edge portion of the first cover 75 fits into the side wall of the main body 60 surrounding the culture chamber 51 and the upper end of the side plate 66. As a result, the first cover 75 is prevented from being displaced in the horizontal direction with respect to the culture chamber 51. At this time, the end portion of the first cover 75 on the sensor chamber 52 side is arranged on the roof portion 65.

10 is a perspective view of the second cover 78, and FIG. 11 is a bottom view of the second cover 78. The second cover 78 is formed in a rectangular plate shape slightly larger than the outer shape of the side wall and the side plate 67 of the main body 60 surrounding the sensor chamber 52 and the adjustment chamber 53, and the outer edge portion is bent downward (right angle). When the sensor chamber 52 and the adjustment chamber 53 are covered with the second cover 78, the outer edge portion of the second cover 78 fits on the side wall of the main body 60 surrounding the sensor chamber 52 and the adjustment chamber 53 and the upper end of the side plate 66. As a result, the second cover 78 is prevented from being displaced in the horizontal direction with respect to the sensor chamber 52 and the adjustment chamber 53. At this time, the end portion of the second cover 78 on the culture chamber 51 side is arranged on the roof portion 65. The procedure for attaching the second cover 78 to the side wall of the main body 60 surrounding the sensor chamber 52 and the adjustment chamber 53 and the upper end of the side plate 66 will be described later.

Through holes 78a and 78b having a circular cross section are formed in a portion of the second cover 78 arranged above the sensor chamber 52.

Cylindrical pins 79A and 79B are provided in the portions of the second cover 78 arranged above the grooves 68A and 68B of the main body 60, respectively. The pins 79A and 79B (first engaging portion) extend horizontally inward from the downwardly folded outer edge portion of the second cover 78. The diameter d1 of the pins 79A and 79B is slightly smaller than the predetermined width W2 of the grooves 68A and 68B. The length L1 of the pins 79A and 79B is substantially equal to the predetermined depth D2 of the grooves 68A and 68B. That is, the pins 79A and 79B can be engaged with the grooves 68A and 68B, respectively.

The second cover 78 is attached to the main body 60 before the first cover 75. The operator attaches the second cover 78, the first cover 75, the oxygen concentration sensor 81, and the pH sensor 82 to the main body 60 by the following procedure.

First, the pins 79A and 79B of the second cover 78 are aligned and moved downward so that the pins 79A and 79B of the second cover 78 engage with the upwardly extending portions of the grooves 68A and 68B, respectively. At this time, the end portion of the second cover 78 on the culture chamber 51 side is located on the roof portion 65. Further, a gap substantially equal to the length of the horizontally extending portions of the grooves 68A and 68B is formed between the side plate 67 and the end portion (downwardly folded outer edge portion) of the second cover 78 on the culture chamber 51 side. ing.

After the pins 79A and 79B reach the lower ends of the upwardly extending portions of the grooves 68A and 68B, respectively, the second cover 78 is horizontally moved away from the culture chamber 51. As a result, the pins 79A and 79B engage with the horizontally extending portions of the grooves 68A and 68B. Therefore, the pins 79A and 79B restrict the movement of the second cover 78 in the vertical direction.

That is, after covering the upper part of the sensor chamber 52 and the adjustment chamber 53 with the second cover 78, the second cover 78 is moved in the direction away from the culture chamber 51, so that the second cover 78 moves in the vertical direction with respect to the main body 60. The grooves 68A and 68B and the pins 79A and 79B are engaged with each other so as to regulate the above.

Subsequently, the position of the first cover 75 is aligned and moved downward so that the outer edge portion of the first cover 75 is fitted to the side wall of the main body 60 surrounding the culture chamber 51 and the upper end of the side plate 66. As a result, the first cover 75 is attached to the main body 60, and the upper part of the culture chamber 51 is covered with the first cover 75. At this time, the gap between the end of the second cover 78 on the culture chamber 51 side (downwardly folded outer edge) and the end of the first cover 75 on the sensor chamber 52 side (downwardly folded outer edge). Is narrower (substantially 0) than the length of the horizontally extending portions of the grooves 68A and 68B. That is, the second cover 78 is moved in the direction closer to the culture chamber 51 by covering the upper part of the culture chamber 51 with the first cover 75 in a state where the grooves 68A and 68B and the pins 79A and 79B are engaged, respectively. Is regulated. As a result, the pins 79A and 79B are restricted from coming off from the grooves 68A and 68B.

Subsequently, as shown in FIG. 12, the sensor cases 81C and 82C are fitted into the through holes 78a and 78b of the second cover 78, respectively. The sensor cases 81C and 82C are formed in a cylindrical shape having an outer diameter slightly smaller than the inner diameters of the through holes 78a and 78b, respectively, and a flange f is provided at the lower portion. The inner diameters of the sensor cases 81C and 82C are slightly larger than the outer diameters of the oxygen concentration sensor 81 and the pH sensor 82, respectively. The sensor cases 81C and 82C are attached to the second cover 78 by inserting the lower ends of the sensor cases 81C and 82C into the through holes 78a and 78b and bringing the flange f into contact with the upper surface of the second cover 78. The second cover 78 and the sensor cases 81C and 82C can also be screwed together.

Subsequently, the oxygen concentration sensor 81 and the pH sensor 82 are inserted into the sensor cases 81C and 82C, respectively. Large diameter portions 81a and 82a having a larger diameter than the other portions are provided above the oxygen concentration sensor 81 and the pH sensor 82. By bringing the lower ends of the large diameter portions 81a and 82a into contact with the upper ends of the sensor cases 81C and 82C, respectively, the oxygen concentration sensor 81 and the pH sensor 82 are attached to the sensor cases 81C and 82C, respectively. The sensor cases 81C and 82C and the oxygen concentration sensor 81 and the pH sensor 82 can also be screwed together. As described above, the second cover 78, the first cover 75, the oxygen concentration sensor 81, and the pH sensor 82 are attached to the main body 60.

Further, as shown in FIGS. 10 and 11, a thin portion 78c having a thickness thinner than the other portions is formed in the portion of the second cover 78 located above the adjustment chamber 53. As a result, when the second cover 78 is attached to the main body 60, the gap between the portion surrounding the adjustment chamber 53 in the main body 60 and the second cover 78 (thin-walled portion 78c) is the portion surrounding the sensor chamber 52 in the main body 60. It is wider than the gap with the second cover 78. That is, the gap between the portion of the main body 60 that surrounds the adjusting chamber 53 and the thin-walled portion 78c serves as a discharge port for discharging gas from the adjusting chamber 53.

The present embodiment described in detail above has the following advantages.

-Inside the culture chamber 51, the cell tissue B is arranged at a predetermined depth D downward from the first opening 55. Then, the culture liquid L is continuously sent from the liquid feeding pipe 42 to the inside of the culture chamber 51 so that the culture liquid L continuously overflows from the first opening 55. In a state where the culture solution L continuously overflows from the first opening 55, the depth of the culture solution L inside the culture chamber 51 is constant. Therefore, the cell tissue B is always arranged at a predetermined depth D from the first opening 55 in the culture medium L. Therefore, the hydraulic pressure of the culture solution L acting on the cell tissue B can be made constant, and the pressure acting on the cell tissue B in the cell culture tank 50 can be stabilized.

A second opening 56 is formed at a position lower than the first opening 55 in the upper part of the sensor chamber 52. The culture solution L overflowing from the first opening 55 of the culture chamber 51 flows into the sensor chamber 52, and the culture solution L overflows from the second opening 56 of the sensor chamber 52. The oxygen concentration sensor 81 and the pH sensor 82 are inserted into the sensor chamber 52, and measure the properties of the culture solution L stored inside the sensor chamber 52. Therefore, the properties of the culture solution L stored inside the sensor chamber 52 can be maintained in the same manner as the properties of the culture solution L stored inside the culture chamber 51, and the properties are changed by the activity of the cell tissue B. The properties of the culture solution L can be measured by the oxygen concentration sensor 81 and the pH sensor 82.

-The culture solution L does not flow from the culture chamber 51 into the sensor chamber 52, and the culture solution L does not flow back from the sensor chamber 52 to the culture chamber 51. Therefore, while making it possible to measure the properties of the culture solution L, it is possible to prevent the culture solution L in the culture chamber 51 from being contaminated when measuring the properties of the culture solution L. Therefore, when the pH of the culture solution L is detected by the pH sensor 82 in the sensor chamber 52, even if the reference solution (internal solution) slightly leaks from the pH sensor 82 to the culture solution L, the culture solution L in the culture chamber 51 Can be prevented from contaminating. Moreover, since the oxygen concentration sensor 81 and the pH sensor 82 are inserted inside the sensor chamber 52, it is possible to prevent the oxygen concentration sensor 81 and the pH sensor 82 from interfering with the maintenance of the culture chamber 51.

The culture chamber 51 and the sensor chamber 52 are adjacent to each other via a first partition wall 61 extending in the vertical direction. The upper end 61a of the first partition wall 61 forms a part of the first opening 55. Therefore, the culture solution L overflowing from the culture chamber 51 over the upper end 61a of the first partition wall 61 flows into the sensor chamber 52. The adjustment chamber 53 is adjacent to the sensor chamber 52 via a second partition wall 62 extending in the vertical direction. The upper end 62a of the second partition wall 62 forms a part of the second opening 56, and the upper end 62a of the second partition wall 62 is lower than the upper end 61a of the first partition wall 61. Therefore, the culture solution L overflowing from the sensor chamber 52 over the upper end 62a of the second partition wall 62 flows into the adjustment chamber 53, and the culture solution L does not flow back from the sensor chamber 52 to the culture chamber 51. Therefore, while making it possible to measure the properties of the culture solution L, it is possible to prevent the culture solution L in the culture chamber 51 from being contaminated when measuring the properties of the culture solution L.

The cell culture tank 50 includes a first cover 75 that covers the upper part of the culture chamber 51, and a second cover 78 that covers the upper part of the sensor chamber 52 and the adjustment chamber 53. Therefore, it is possible to prevent germs and foreign substances from entering the culture chamber 51, the sensor chamber 52, and the adjustment chamber 53. Further, in the configuration in which the oxygen concentration sensor 81 and the pH sensor 82 penetrate the second cover 78 and are inserted into the sensor chamber 52, only the first cover 75 can be opened and closed for maintenance.

The cell culture tank 50 is provided with a roof portion 65 that covers the upper part of the first partition wall 61. The end of the first cover 75 on the sensor chamber 52 side and the end of the second cover 78 on the culture chamber 51 side are arranged on the roof portion 65. Therefore, even if bacteria or foreign substances fall from the gap between the first cover 75 and the second cover 78, they can be received by the roof portion 65, and the bacteria and foreign substances are suppressed from entering the culture chamber 51 and the sensor chamber 52. can do.

The cell culture tank 50 includes a main body 60 that surrounds the sensor chamber 52 and the adjustment chamber 53 together with the first partition wall 61. That is, the sensor chamber 52 and the adjustment chamber 53 are surrounded by the first partition wall 61 and the main body 60. The second cover 78 and the main body 60 have a second cover 78 with respect to the main body 60 by covering the upper part of the sensor chamber 52 and the adjustment chamber 53 with the second cover 78 and then moving the second cover 78 in a direction away from the culture chamber 51. 2 Pins 79A and 79B and grooves 68A and 68B that engage with each other so as to restrict the vertical movement of the cover 78 are provided, respectively. Therefore, by engaging the pins 79A and 79B with the grooves 68A and 68B, respectively, it is possible to restrict the vertical movement of the second cover 78 with respect to the main body 60, and the second cover 78 unintentionally opens. Can be suppressed.

-By covering the upper part of the culture chamber 51 with the first cover 75 in a state where the pins 79A and 79B and the grooves 68A and 68B are engaged, the second cover 78 can be moved in the direction closer to the culture chamber 51. It is regulated. Therefore, by covering the upper part of the culture chamber 51 with the first cover 75, it is possible to prevent the pins 79A and 79B from being disengaged from the grooves 68A and 68B, and the second cover 78 is unintentionally opened. That can be further suppressed.

The first cover 75 is formed with an introduction port 76 for introducing gas into the inside of the culture chamber 51. Therefore, a gas for adjusting the properties of the culture solution L stored in the culture chamber 51 can be introduced into the inside of the culture chamber 51 from the introduction port 76. Further, the gas introduced into the inside of the culture chamber 51 from the introduction port 76 is introduced into the sensor chamber 52 through the first opening 55, and is introduced into the adjustment chamber 53 through the second opening 56. Therefore, the properties of the culture solution L in the sensor chamber 52 can be brought close to the properties of the culture solution L in the culture chamber 51.

The first cover 75 is formed with an introduction port 76 for introducing gas into the inside of the culture chamber 51, and the second cover 78 has a portion of the main body 60 surrounding the adjustment chamber 53 and the second cover 78. A thin portion 78c is formed so that the gap is wider than the gap between the portion of the main body 60 surrounding the sensor chamber 52 and the second cover 78. Therefore, the gas introduced into the adjusting chamber 53 is easily discharged to the outside of the adjusting chamber 53 through the gap between the main body 60 and the thin portion 78c, and the culture chamber 51 goes to the sensor chamber 52 and the sensor chamber 52 goes to the adjusting chamber 53. And in order, it becomes easier to guide the gas. Therefore, the properties of the culture solution L in the sensor chamber 52 can be further brought closer to the properties of the culture solution L in the culture chamber 51.

The inside of the airtight tank 34 by adjusting the flow rate Q1 of the air supplied from the air supply source 21 by the flow rate proportional valve 24 and limiting the flow rate Qr of the air discharged to the discharge unit 28 by the throttle valve 27. Pressure Pv can be controlled. Then, the cell culture tank 50 stores the culture solution L and the cell tissue B. The liquid feeding pipe 42 connects the inside of the culture liquid L stored in the tank 34 to the cell culture tank 50. Therefore, by supplying air to the tank 34 and increasing the pressure Pv inside the tank 34, the culture solution L is sent from the tank 34 to the cell tissue B of the cell culture tank 50 via the liquid supply pipe 42. be able to.

When the culture liquid L is sent from the tank 34 to the cell culture tank 50 via the liquid supply pipe 42, the control unit 45 supplies the culture liquid L from the air supply source 21 in a state where the pressure Pv inside the tank 34 is balanced. The flow rate proportional valve 24 is controlled so that the target flow rate of air corresponding to the target pressure inside the tank 34 is supplied based on a predetermined correlation between the flow rate Q1 of the air to be generated and the pressure Pv inside the tank 34. In this state, the pressure Pv inside the tank 34 is balanced. Therefore, the culture solution L can be sent to the cell culture tank 50 in a state where the pressure Pv inside the tank 34 is equilibrated to the target pressure, as compared with the case where the culture solution L of the target flow rate is sent. , The culture solution L can be sent to the cell tissue B of the cell culture tank 50 in a state closer to the living body.

When the pressure Pv inside the tank 34 is in equilibrium, the pressure Pv inside the tank 34 and the air flow rate Q1 supplied from the air supply source 21 have a correlation with a small influence of temperature. .. Therefore, it is possible to suppress the feeding state of the culture solution L from being affected by disturbance due to the ambient temperature.

The target pressure is 10 [mmHg] or less, and the time from the start of supplying air from the air supply source 21 to the equilibration of the pressure Pv inside the tank 34 is 5 [S] or less. According to such a configuration, the pressure Pv inside the tank 34 can be equilibrated in a short time of 5 [S] or less, and the culture solution L can be quickly sent to the cell culture tank 50 at the target pressure.

The cell culture system 10 includes a pump 90 for pumping the culture solution L, and a return pipe 43 connecting the tank 34 and the pump 90. Therefore, the culture solution L can be replenished to the tank 34 via the return liquid pipe 43.

The liquid return pipe 43 is connected to the tank 34 at a position lower than the liquid level of the culture liquid L and higher than the lower end of the liquid feed pipe 42. Therefore, as compared with the case where the return liquid pipe 43 is connected to the tank 34 at a position higher than the liquid level of the culture liquid L, it is possible to suppress the culture liquid L from falling to the liquid surface and generating bubbles. be able to.

-Suppressing that air bubbles mixed in the culture solution L are taken into the liquid feed pipe 42 as compared with the case where the liquid return pipe 43 is connected to the tank 34 at a position lower than the lower end of the liquid feed pipe 42. Can be done.

A check valve 92 is provided in the return pipe 43 to allow the flow of the culture solution L from the pump 90 to the tank 34 and prohibit the flow of the culture solution L from the tank 34 to the pump 90. Therefore, even if the return liquid pipe 43 is connected to the tank 34 at a position lower than the liquid level of the culture liquid L and the pressure of the culture liquid L is applied to the return liquid pipe 43, the culture liquid L is applied from the tank 34 to the pump 90. Can be suppressed from flowing back.

It should be noted that the above embodiment can be modified and implemented as follows. The same parts as those in the above embodiment are designated by the same reference numerals, and the description thereof will be omitted.

-The thin portion 78c can be omitted in the second cover 78. Even in that case, the gas can be discharged from the gap between the main body 60 and the second cover 78.

If it is not necessary to adjust the dissolved oxygen concentration DO and pH of the culture solution L, the introduction port 76 may be omitted in the first cover 75.

Pins 79A and 79B may be provided on the main body 60, and grooves 68A and 68B may be provided on the second cover 78. In that case, the grooves 68A and 68B may be extended upward and then horizontally in the direction of the culture chamber 51. Further, the grooves 68A and 68B of the main body 60 and the pins 79A and 79B of the second cover 78 can be omitted.

-The first cover 75 and the second cover 78 may be integrally formed. Further, when the main body 60 is arranged inside the incubator or the like, the first cover 75 and the second cover 78 may be omitted. In that case, the roof portion 65 may also be omitted.

As shown in FIG. 13, the height of the upper end of the first partition wall 161 may be changed between the first upper end 161a and the second upper end 161b. In the first partition wall 161, a second upper end 161b is provided in a portion adjacent to the adjustment chamber 53, and a first upper end 161a is provided in the remaining portion. The second upper end 161b is higher than the first upper end 161a. According to such a configuration, it is possible to prevent the culture solution L overflowing from the culture chamber 51 from directly flowing into the adjustment chamber 53. Therefore, the culture solution L can be reliably flowed from the culture chamber 51 to the sensor chamber 52 and the adjustment chamber 53 in this order. It is also possible to extend the second upper end 161b to the bottom of the roof portion 65 so that the first opening 55 is not formed above the second upper end 161b.

The culture solution L can be sent from the liquid feeding pipe 42 to the inside of the culture chamber 51 in a predetermined cycle T so that the culture solution L overflows from the first opening 55 in a predetermined cycle T. The predetermined cycle T is a cycle in which evaporation of the culture solution L does not matter, and for example, 1 second to 1 minute can be adopted. In the state where the culture solution L overflows from the first opening 55 or immediately after the overflow, the depth of the culture solution L inside the culture chamber 51 is constant. Therefore, the cell tissue B is arranged at a predetermined depth D from the first opening 55 in the culture medium L. Therefore, the hydraulic pressure of the culture solution L acting on the cell tissue B can be made constant, and the pressure acting on the cell tissue B in the cell culture tank 50 can be stabilized. The predetermined cycle T can also be made variable according to the air temperature (evaporation rate of the culture solution L).

-It is also possible to omit the adjustment chamber 53 and connect the liquid return pipe 43 to the sensor chamber 52.

-If it is not necessary to detect the properties of the culture solution L, the oxygen concentration sensor 81 and the pH sensor 82 can be omitted. Further, if it is not necessary to detect the properties of the culture solution L, the sensor chamber 52, the oxygen concentration sensor 81, and the pH sensor 82 can be omitted. In that case, the culture solution L overflowing from the culture chamber 51 can be discarded.

-Instead of the flow rate proportional valve 24, an open / close solenoid valve is adopted, and the control unit 45 can also adjust the flow rate of air by controlling the valve opening ratio of the solenoid valve in a predetermined period by DUTY.

The throttle valve 27 (throttle portion) is not limited to the needle valve, and may be a fixed orifice.

The return liquid pipe 43 can be connected to the tank 34 at a position lower than the liquid level of the culture liquid L and lower than the lower end of the liquid feed pipe 42. Further, the liquid return pipe 43 can be connected to the tank 34 at a position higher than the liquid level of the culture liquid L.

-Animal cells, plant cells, insect cells, bacteria, yeasts, fungi and the like can be adopted as the cell tissue B or in place of the cell tissue B. Further, the cell tissue B may be a tissue composed of a plurality of types of cells.

-Depending on the type of cell tissue B (cell), the target pressure is higher than 10 [mmHg] and 20 [mmHg] or less, and after starting to supply air from the air supply source 21, the inside of the tank 34 It is also possible to adopt a configuration in which the time until the pressure Pv is equilibrated is longer than 5 [S] and shorter than 10 [S]. Further, the target pressure is higher than 20 [mmHg], and the time from the start of supplying air from the air supply source 21 to the equilibration of the pressure Pv inside the tank 34 is longer than 10 [S]. A configuration can also be adopted.

-It is also possible to adopt a supply source that supplies nitrogen (gas) instead of the air supply source 21.

10 ... Cell culture system, 20 ... Pressure control liquid supply unit, 34 ... Tank, 42 ... Liquid supply pipe, 43 ... Liquid return pipe, 50 ... Cell culture tank, 51 ... Culture room (1st room), 52 ... Sensor room (2nd room), 53 ... Adjustment room (3rd room), 55 ... 1st opening, 56 ... 2nd opening, 60 ... Main body, 61 ... 1st partition wall (1st wall), 62 ... 2nd partition wall (2nd wall), 75 ... 1st cover (1st lid), 78 ... 2nd cover (2nd lid), 81 ... Oxygen concentration sensor (sensor), 82 ... pH sensor (sensor), 161 ... 1st partition Wall (first wall).

Claims (8)

The first room with the first opening formed at the top,
A liquid supply pipe that sends the culture solution to the inside of the first chamber,
Inside the first chamber, a cell tissue arranged at a predetermined depth downward from the first opening,
Equipped with
A cell culture tank in which the culture solution is continuously fed from the liquid feeding pipe to the inside of the first chamber so that the culture solution continuously overflows from the first opening. The first room with the first opening formed at the top,
A liquid supply pipe that sends the culture solution to the inside of the first chamber,
Inside the first chamber, a cell tissue arranged at a predetermined depth downward from the first opening,
Equipped with
A cell culture tank in which the culture solution is sent from the liquid feeding pipe to the inside of the first chamber in the predetermined cycle so that the culture solution overflows from the first opening in the predetermined cycle. A second opening is formed in the upper part at a position lower than the first opening, and the culture solution overflowing from the first opening flows into the second chamber, and the culture solution overflows from the second opening.
A sensor inserted into the second chamber and measuring the properties of the culture solution stored inside the second chamber.
The cell culture tank according to claim 1 or 2. The first chamber and the second chamber are adjacent to each other via a first wall extending in the vertical direction.
The upper end of the first wall forms a part of the first opening.
A third room adjacent to the second room is provided via a second wall extending in the vertical direction.
The cell culture tank according to claim 3, wherein the upper end of the second wall forms a part of the second opening, and the upper end of the second wall is lower than the upper end of the first wall. The first lid that covers the upper part of the first chamber and
A second lid that covers the second chamber and the third chamber, and
The roof covering the upper part of the first wall and
Equipped with
The sensor is inserted into the inside of the second chamber through the second lid.
The cell culture tank according to claim 4, wherein the end portion of the first lid on the second chamber side and the end portion of the second lid on the first chamber side are arranged on the roof portion. A main body that surrounds the second and third chambers together with the first wall is provided.
The second lid and the main body are covered with the second lid above the second chamber and the third chamber, and then the second lid is moved in a direction away from the first chamber. A first engaging portion and a second engaging portion that engage with each other are provided so as to restrict the vertical movement of the second lid with respect to the main body, respectively.
By covering the upper part of the first chamber with the first lid in a state where the first engaging portion and the second engaging portion are engaged, the second lid is brought closer to the first chamber. The cell culture tank according to claim 5, wherein the cell culture tank is regulated to move. The cell culture tank according to claim 5 or 6, wherein the first lid is formed with an introduction port for introducing gas into the first chamber. The first lid is formed with an introduction port for introducing gas into the inside of the first chamber.
The second lid has a thin wall in which the gap between the portion of the main body surrounding the third chamber and the second lid is wider than the gap between the portion of the main body surrounding the second chamber and the second lid. The cell culture tank according to claim 6, wherein the portion is formed.

Images